PROGESTERONE PRODUCTION BY DISSOCIATED RAT OVARIAN CELLS: A SENSITIVE METHOD FOR QUANTITATION OF HORMONES WITH LUTEINIZING ACTIVITY¹

Abstract

A simple method has been developed for the preparation of dissociated rat ovarian cells, capable of responding to luteinizing hormone (LH) or human chorionic gonadotropin (hCG) with production of progesterone. The method involves agitation and trituration of quartered rat ovaries in collagenase-containing medium. Over 80% of the dissociated cells were intact and viable, and remained responsive for at least 48 hours of storage at 4οC. The minimum detectable quantity of hCG was approximately 3 mIU/ml medium, and a linear relationship between log dose and arithmetic response was observed between 3 and 30 mlU with an average index of precision (λ) of 0.043. Similar dose response curves were obtained with highly purified hLH. In contrast, highly purified follicle stimulating hormone (hFSH) in amounts up to 40ng/tube failed to stimulate progesterone production.